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US-12616720-B2 - Cell preparation for extemporaneous use, useful for healing and rejuvenation in vivo

US12616720B2US 12616720 B2US12616720 B2US 12616720B2US-12616720-B2

Abstract

The present invention relates to new plasma or new platelet-rich plasma preparations, new cell dissociation methods, new cell associations or compositions, a method of preparation thereof, a use thereof, devices for the preparation thereof and preparations containing such a platelet-rich plasma preparation and cell associations or compositions. Specifically, the invention provides plasma or platelet-rich plasma alone or in cell composition preparations for use in tissue regeneration and bone regeneration and pain reduction.

Inventors

  • Antoine Turzi
  • Donald DU TOIT

Assignees

  • REGENLAB USA LLC

Dates

Publication Date
20260505
Application Date
20220610
Priority Date
20060821

Claims (20)

  1. 1 . A non-activated cosmetic composition comprising: a platelet concentrate wherein the platelet concentrate is obtained by a one-step centrifugation process that separates erythrocytes while retaining plasma fractions enriched in functionally active platelets, the process comprising: introducing whole blood to a separator tube comprising an anticoagulant composition and a thixotropic gel; centrifuging the separator tube at a force between about 1500 g and about 2000 g to separate plasma from red blood cells of the whole blood; collecting a plasma fraction enriched in platelets without removal of platelet poor plasma resulting in retention of plasma proteins; and re-suspending platelets in remaining plasma after optionally discarding a portion of platelet poor plasma supernatant; wherein the platelet concentrate: concentrates plasma in leucocytes, thrombocytes, and adhesion proteins; comprises at least 300 billion platelets per liter; exhibits 2-4 times platelet and growth factor levels compared to a natural blood clot; maintains fibrin and fibrinogen levels comparable to the natural blood clot; and maintains stability of platelet-derived growth factors for at least 48 hours post-preparation at room temperature.
  2. 2 . The non-activated cosmetic composition according to claim 1 , wherein the platelet concentrate contains greater than 350 billion platelets per liter.
  3. 3 . The non-activated cosmetic composition according to claim 1 , wherein the platelet concentrate contains greater than 400 billion platelets per liter.
  4. 4 . The non-activated cosmetic composition according to claim 1 , wherein the composition is in a form selected from the group consisting of a cream, emulsion, hydrogel, a readily syringable composition, and combinations thereof.
  5. 5 . The non-activated cosmetic composition according to claim 4 , wherein the cosmetic composition is a wrinkle filler composition.
  6. 6 . A method for promoting skin regeneration in a scar or wrinkle comprising applying the non-activated cosmetic composition of claim 1 to a scar or wrinkle.
  7. 7 . The method of claim 6 , wherein applying the non-activated cosmetic composition to a scar or wrinkle comprises filling a wrinkle line with the non-activated cosmetic composition.
  8. 8 . The method according to claim 6 , wherein applying the non-activated cosmetic composition to a scar or wrinkle comprises administering the non-activated cosmetic composition subcutaneously.
  9. 9 . The method according to claim 6 , wherein the platelet concentrate of the non-activated cosmetic composition contains greater than 350 billion platelets per liter.
  10. 10 . The method according to claim 9 , wherein the platelet concentrate of the non-activated cosmetic composition contains greater than 400 billion platelets per liter.
  11. 11 . A method of making the non-activated cosmetic composition of claim 1 , the method comprising: forming plasma containing a platelet concentrate by: introducing whole blood to a separator tube comprising an anticoagulant composition and a thixotropic gel; centrifuging the separator tube only once to separate plasma containing the platelet concentrate from red blood cells of the whole blood; resuspending the plasma without supernatant removal; and collecting the platelet concentrate from the separator tube, wherein the collected platelet concentrate is in a final form for injection into a subject without admixing with an activator, and wherein the collected platelet concentrate contains platelet poor plasma and platelet rich plasma, thereby making the non-activated cosmetic composition of claim 1 .
  12. 12 . The method according to claim 11 , wherein the separator tube is centrifuged at 3,800 revolutions per minute (rpm) for 3.5 minutes.
  13. 13 . The method according to claim 11 , wherein the anticoagulant is present in the separator tube an amount of 3.5 mg/mL.
  14. 14 . The method according to claim 11 , wherein the separator tube is optionally coated.
  15. 15 . The method according to claim 11 , wherein the platelet concentrate of the cosmetic composition contains greater than 350 billion platelets per liter.
  16. 16 . The method according to claim 15 , wherein the platelet concentrate of the cosmetic composition contains greater than 400 billion platelets per liter.
  17. 17 . The non-activated cosmetic composition of claim 1 , comprising mixing the separator tube after centrifuging.
  18. 18 . The non-activated cosmetic composition of claim 17 , wherein mixing comprises inverting the separator tube several times to mix the plasma.
  19. 19 . The non-activated cosmetic composition of claim 1 , wherein collecting the plasma comprises collecting the entire plasma separated from the red blood cells, without removal of platelet poor plasma.
  20. 20 . A method for promoting skin regeneration in a scar or wrinkle comprising applying the non-activated cosmetic composition of claim 19 to a scar or wrinkle.

Description

CROSS-REFERENCE TO RELATED APPLICATIONS This application is a Continuation of U.S. Ser. No. 17/370,865, filed Jul. 8, 2021, which is a Continuation of U.S. Ser. No. 17/065,098, filed Oct. 7, 2020, now U.S. Pat. No. 11,241,458, which is a Continuation of Ser. No. 16/793,296, filed Feb. 18, 2020, now U.S. U.S. Pat. No. 11,096,966, which is a Continuation of U.S. Ser. No. 16/103,453, filed Aug. 14, 2018, now U.S. Pat. No. 10,881,691, which is a Continuation of U.S. Ser. No. 15/605,696, filed May 25, 2017, now U.S. Pat. No. 10,052,349, which is a Continuation of U.S. Ser. No. 15/044,498, filed Feb. 16, 2016, now U.S. Pat. No. 10,092,598, which is a Continuation of U.S. Ser. No. 14/021,196, filed Sep. 9, 2013, now abandoned, which is a Continuation of U.S. Ser. No. 12/438,236, filed Feb. 20, 2009, now U.S. Pat. No. 8,529,957, which is the U.S. national phase application of International Patent Application No. PCT/EP2007/058695, filed Aug. 21, 2007, which claims priority to PCT/EP2006/065493, filed Aug. 21, 2006, the disclosures of each of which are hereby incorporated by reference in their entirely, including all figures, tables and amino acid or nucleic acid sequences. TECHNICAL FIELD The present invention is related to the field of tissue regeneration, especially skin, cartilage, muscle, tendon, adipose tissue, cornea, peripheral nerves, spine and bone regeneration. It concerns more particularly new cell preparations as a biological scaffold, a method of preparation thereof, a use thereof, a device for the preparation thereof and preparations containing such cell preparation for extemporaneous use. BACKGROUND The importance of biological autologous materials in the healing process has been well documented. Most importantly, two biological autologous materials have been shown to be directly implicated in the formation of the structure of blood clots, which provide a haemostatic barrier whose role is to ensure hemostasis and seal the wound: (1) fibrin, which derives from the separation of plasma fibrinogen into two strands through the action of thrombin, and (2) the activated membranes of platelets. The wound healing process is generally presented as the succession of a coagulation phase, an inflammatory process and a regeneration process. The coagulation phase (blood clotting or clot formation) is a complex process whereby a damaged blood vessel wall is covered by a fibrin clot to stop hemorrhage and the repair of the damaged vessel is initiated by the release in large quantities of cytokines and growth factors from platelet alpha granules. The formation of blood clots (formed in physiological conditions by fibrin, platelets and red blood cells, among other blood components) is a natural phenomenon that results from tissue trauma and its role in the wound healing process, as well as in the union of bone fractures, is well-known. The inflammation process, which follows the formation of a blood clot, is stimulated by numerous vasoactive mediators and chemotactic factors (specific signals in the form of proteins) released by white blood cells and platelets. These signals attract macrophages that “clean” the site from bacteria and foreign particles as well as red blood cells before the migration of new cells. The tissue regeneration phase involves the chemoattraction and the mitosis of the undifferentiated cells in the scaffold (or growth matrix) formed by the blood clot. The new cells which multiply under the stimulation of platelet growth factors will replace damaged or destroyed cells injured by macrophages. Growth factors and numerous plasma proteins, also called signaling molecules, which promote cell migration and division within blood clots, play a crucial role in the wound healing process. Theoretically, it is possible to amplify the effects of these first phases in the wound-healing cascade by discarding the red blood cells and increasing the concentration of growth factors. Blood clotting amplification can be defined as the formation of an “enriched clot (EC)”. ECs are obtained through the use of platelet concentrates and have been described in Platelets and Megacaryocytes 2004, vol 1 & 2, as “Structure and signals”, Ed. Gibbins and Mahaut-Smith, Humana Press, New Jersey. Platelet-rich plasma (PRP) can be defined as an autologous concentrate of platelets in a small volume of plasma; it has been developed as an autologous biomaterial and has proven to be useful in the healing and regeneration of tissues (Marx et al., 2004, J. Oral Maxillofac. Surg., 62, 489-496). PRP not only consists in a platelet concentrate but also contains growth factors (such as platelet-derived growth factor: PDGF, vascular endothelial growth factor: VEGF, transforming growth factor: TGF and epidermal growth factor: EGF) that are actively secreted by platelets and are known to have a fundamental role in wound healing initiation process. For example, PDGF is known to initiate connective tissue healing, including bone regeneration and